Effect of Fufang Huangbai Fluid Paint on Virulence and Biofilm of Methicillin-resistant Staphylococcus aureus
10.13422/j.cnki.syfjx.20220609
- VernacularTitle:复方黄柏液涂剂对耐甲氧西林金黄色葡萄球菌毒力和生物膜的影响
- Author:
Jin-ze LI
1
;
Kai-yu CUI
2
;
Dong-ying LI
2
;
Shu-hua MA
2
;
Gai-ying HE
2
;
Ya-nan SUN
2
;
Yi WANG
2
;
Zhong-mei HE
1
;
Wei-feng YANG
2
Author Information
1. Jilin Agricultural University，Changchun 130118，China
2. Experimental Research Center，China Academy of Chinese Medical Sciences，Beijing 100700，China
- Publication Type:Journal Article
- Keywords:
Fufang Huangbai Fluid Paint （FFHBFP）;
methicillin-resistant Staphylococcus aureus;
virulence factor;
biofilm
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(8):54-62
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo study the virulence and biofilm inhibition effect of Fufang Huangbai Fluid Paint （FFHBFP） on methicillin-resistant Staphylococcus aureus （MRSA）， and to explore the antibacterial effect of FFHBFP on MRSA， which provides a theoretical basis and reference for clinical medication. MethodFirstly， the microdilution method and time–growth curve were used to determine the minimum inhibitory concentration （MIC） of FFHBFP and vancomycin （VAN） against MRSA and the effect on bacterial growth. The effects of FFHBFP and VAN on the inhibition of MRSA virulence factor lipase and restoration of hydrogen peroxide （H2O2） sensitivity were detected under sub-minimum inhibitory concentration （sub-MIC）. The inhibitory effect of FFHBFP and VAN on MRSA biofilm formation and maturation was detected by the microplate method. The morphological changes of mature biofilms before and after administration were observed under a scanning electron microscope （SEM）. Real-time polymerase chain reaction （Real-time PCR） was utilized to detect the effect of 50.600 g·L-1 concentration of FFHBFP on the expression of MRSA virulence gene crtM and biofilm-forming genes fnbA and icaA. Finally， molecular docking technology was used to predict the mechanism of potential antibacterial active ingredients of FFHBFP in inhibiting the virulence and biofilm of MRSA. ResultThe MIC of VAN was 2 mg·L-1， and VAN below 1 mg·L-1 exerted no effect on MRSA growth. The MIC of FFHBFP was not determined， while the 101.200-202.400 g·L-1 original solution inhibited MRSA growth. Compared with the blank group and the VAN group， sub-MIC （25.300-50.600 g·L-1 original solution） inhibited lipase and recovered MRSA sensitivity to H2O2 （P<0.01）. The results of the microplate method showed that FFHBFP （25.300-202.400 g·L-1 original solution） inhibited biofilm formation and maturation （P<0.05， P<0.01）. The SEM exhibited that FFHBFP made the structure of biofilm loose and the size of the bacteria varied. FFHBFP at 50.600 g·L-1 concentration can inhibit the expression of related virulence genes and biofilm-forming genes （P<0.05， P<0.01）， and molecular docking results also showed that the main antibacterial active ingredients in FFHBFP have good binding ability to the target. ConclusionFFHBFP that cannot directly kill MRSA exerts clinical efficacy by impairing virulence expression， biofilm formation， and other pathogenic properties.
